SERUM

A simple, fast and sensitive quantification method for the drug metformin in dog serum was developed using flow injection analysis (FIA)-tandem mass spectrometry (MS/MS). The method was fully validated according to industry standards. It is the first time that FIA-MS/MS for metformin was developed surpassing all existing methods in terms of time of analysis. The quantification method was dependent on the formation of [M+H](+) using electrospray ionization (ESI) and employing multiple reaction monitoring (MRM) using quadrupole-linear ion trap (4000 QTRAP(®)) instrument. A deuterated internal standard (IS) of metformin bearing six deuterium atoms was used to compensate for matrix effects and for variation in ion current within the ESI source. The ion transitions that were monitored were m/z 130.1→m/z 71.0 and m/z 130.1→m/z 60.1 for metformin and m/z 136.0→m/z 77.0 for the internal standard. A linear response (r=0.9966) was established for a range of concentrations of 5-2340ng/mL. The ...

ABSTRACT
An experiment was conducted to study the effect of dietary vitamin E on performance, digestibility and hematological parameters for a period of eight weeks with 96 day-old marshal broiler chicks. Basal diets was supplemented with different levels of vitamin E which are 0.5g/kg, 1.0g/kg and 1.5g/kg. Each test diet was fed for 8 weeks ad libitum in starter and finisher form. Result of the present study shows significant difference (p<0.05) in body weight gain with highest and lowest mean values are (712.60g and 653.36g) respectively. Whereas feed intake showed significant difference (p<0.05).
Haematological parameters revealed that PCV is affected by varying levels of vitamin E supplementation, also differential counts lymphocytes, monocytes and heterophils showed significant difference (p<0.05) across the treatments. Haemoglobin decreased with increasing level of inclusion of vitamin E supplementation. Serum result from present study shows that only LDL is significantly different (p<0.05) across the treatment while serum cholesterol increase with increasing level of supplementation with treatment 3 having the highest mean value of 113.07mg/dl and treatment 1 with the lowest at 1005.56md/dl. Digestibility of nutrients result of present study shows that there are no significant difference in dry matter digestibility as digestibility was fair across all treatments, also crude protein digestibility was highest in treatment 3 with mean value of 65.84±2.76and lowest in the control diet with mean value of 56.38±7.04. Varying levels of vitamin E did not significantly change the ether extract values.
KEY WORDS: Vitamin E, Broilers, Haematological parameters, Digestibility, Performance, Serum chemistry.

Chikungunya fever is a mosquito-borne disease of key public health importance in tropical and subtropical countries. Although severe joint pain is the most distinguishing feature of chikungunya fever, diagnosis remains difficult because the symptoms of chikungunya fever are shared by many pathogens, including dengue fever. The present study aimed to develop a new immunochromatographic diagnosis test for the detection of chikungunya virus antigen in serum. Mice were immunized with isolates from patients with Thai chikungunya fever, East/Central/South African genotype, to produce mouse monoclonal antibodies against chikungunya virus. Using these monoclonal antibodies, a new diagnostic test was developed and evaluated for the detection of chikungunya virus. The newly developed diagnostic test reacted with not only the East/Central/South African genotype but also with the Asian and West African genotypes of chikungunya virus. Testing of sera from patients suspected to have chikungunya f...

The purpose of this study was to isolate and identify potential lactic acid bacteria from palm raffia wine and to evaluate in vivo the effect of isolated probiotic bacteria on serum lipids and some enzymes activities in wistar albino rats. Lactic acid bacteria were isolated on MRS agar using pour plate method. The catalase negative and Gram positive isolates were selected as presumptive lactic acid bacteria. They were also assessed for invitro cholesterol assimilation from culture media, acid tolerance and bile salt tolerance. Selected isolates were biochemically characterized using the API 50 CHL BioMerieux kit. In-vivo cholesterol lowing effect was studied on hyperlipidemic induced wistar rats randomly assigned into three groups. The experiment was carried out for four weeks by oral gavages; dose level 10 8-10 10 CFU/ ml. The administered volume of each dose was 1.0 ml/kg body weight/ day, after the feeding trial, the rats were dissected and blood for serum collected for lipid profile (triglyceride, total cholesterol, LDL-c, VLDL-c and HDL-c), transaminases (ALT and AST) analysis using kits. 35 colonies from palm were studied and 22 were Gram positive. Catalase negative text allowed the selection 10 of them which were studied for their acid and bile tolerance; Data from acid and bile tolerance assays showed that seven isolates were able to maintain viability at pH 2 and to grow in a medium with 0.4% of bile salts. After in vitro assessment for their cholesterol-lowering action 3 isolates were finally selected and identified using the API 50 CHL as two L. plantarum and one L. pentosus. These strains gave rise to a significant reduction (P < 0.05) of the cholesterol level in MRS broth. In particular, L. pentosus strain lowered the cholesterol content by an average of 76.43%, compared to L. plantarum strains which lowered cholesterol content by an average of 51.25% for the isolate 2R30 and 55.22% for 2R36 repectively. The results showed no adverse effect on rats organs according to the enzyme activities and no sign of toxicity also. The probiotic strains (Lactobacillus pentosus) selected effectively demonstrated their effect on reduction of the levels of total cholesterol, LDL and VLDL in the rats blood serum. Data from acid and bile tolerance, in-vitro cholesterol reduction and in-vivo lipidemia assays showed that the dairy isolate L. pentosus can therefore be regarded as new probiotic for and could be used for functional food production.

Objectives. To evaluate and compare the performances of three enzyme-immunoassays (EIAs) and a double radial immunodiffusion (DRID) test in addition to immunofluorescence (IF) microscopy for routine laboratory screening of patient sera sent for antinuclear antibody (ANA) analysis.Methods. 3079 consecutive patient sera sent for routine testing of ANA were analysed by IF microscopy on HEp-2 cells (IF-ANA), three different ANA-EIAs, and a DRID test for antibodies against extractable nuclear antigens. The IF-ANA and DRID tests were regarded as reference methods.Results. By IF-ANA and/or DRID, 375 sera (12%) turned out ANA-positive. A further 171 sera (6%) were positive by EIA, but could not be confirmed either by IF microscopy or DRID. 32 of the 375 ANA-positive (9%) sera were negative by IF microscopy, but had precipitating antibodies against Ro/SS-A (52 and/or 60 kD).Conclusions. Different assays for ANA analysis give overlapping results to a certain extent, but are by no means interchangeable. Thus, different ANA tests reflect different aspects of these autoantibodies. The diagnostic utility of ANA testing still mainly refers to IF-microscopy and precipitin tests. IF-ANA should not be abandoned as the golden standard in clinical routine, until diagnostic and classification criteria for systemic lupus erythematosus and other systemic inflammatory autoimmune diseases have been revised. However, in addition we strongly advocate that a specific test for anti-Ro/SS-A antibodies is always included.

Fabrication of an amperometric cholesterol biosensor by co-immobilization of cholesterol esterase (ChEt) and cholesterol oxidase (ChOx) onto conducting polypyrrole (PPY) films using electrochemical entrapment technique is described. Electrochemical polymerization was carried out using a two-electrode cell configuration at 0.8 V. Characterization of resulting amperometric biosensor for the estimation of cholesterol has been experimentally determined in terms of linear response range, optimum pH, applied potential, temperature, and shelf-life. These PPY/ChEt/ChOx electrodes can be used for cholesterol ester estimation from 1 to 8 mM and have shelf-life of about 4 weeks at 4 °C during which about 15 estimations of cholesterol ester could be made. The sensitivity of PPY/ChEt/ChOx electrode has been found to be 0.15 μA/mM and the apparent Km value for this electrode is 9.8 mM. Conductivity of the polymer films found to be about 3×10−3 S/cm.

It has been known for over 20 years that foetal calf serum can induce hypertrophy in cultured cardiomyocytes but this is rarely considered when examining cardiomyocytes derived from pluripotent stem cells (PSC). Here, we determined how serum affected cardiomyocytes from human embryonic- (hESC) and induced pluripotent stem cells (hiPSC) and hiPSC from patients with hypertrophic cardiomyopathy linked to a mutation in the MYBPC3 gene. We first confirmed previously published hypertrophic effects of serum on cultured neonatal rat cardiomyocytes demonstrated as increased cell surface area and beating frequency. We then found that serum increased the cell surface area of hESC- and hiPSC-derived cardiomyocytes and their spontaneous contraction rate. Phenylephrine, which normally induces cardiac hypertrophy, had no additional effects under serum conditions. Likewise, hiPSC-derived cardiomyocytes from three MYBPC3 patients which had a greater surface area than controls in the absence of serum...